A normalized model based on Taqman real-time PCR assay for quantitative comparison of chicken adulteration in raw and heat-treated hamburgers

Mislabeling of hamburgers with undeclared chicken has frequently been reported. A fast TaqMan real-time PCR assay was developed to estimate chicken percentage in raw and heat-treated hamburgers in the current work. The calibration curve was plotted with the normalization approach (ΔCt; the difference calculation between chicken-cytb and eukaryotic-18S rRNA assays) for five reference chicken mixtures (100-0.01%) using a duplex TaqMan-qPCR protocol. The R 2 and efficiency values of the normalized curve were 0.9993 and ~100%, respectively. In the validation plan, analysis of model samples (raw, oven-cooked and autoclaved) under repeatability conditions represented the bias values within the range of ±25% and RSD values <25%, except for a few autoclaved model samples. Therefore, this developed and validated model could be considered a useful and reliable method for estimating chicken species (with a sensitivity of 0.01%) in raw and cooked hamburgers.